تفاعل #7437
ord-34edcaa7e3ba4173928e329654c5e7d9
معادلة التفاعل
bicarbonate
sulfo-NHS-LC-biotin
→
Biotin
المتفاعلات
الكواشف
لا شيء
المذيبات
ظروف التفاعل
الظروف التفصيلية
See reaction.notes.procedure_details.
المعالجة
- 1أخرىto give a final volume of 1.0 mL
- 2أخرىto give a 1 mg/mL solution just
- 3workup.ADDITIONto addition
- 4workup.ADDITIONThe first aliquot (10 uL) is added
- 5workup.ADDITIONThe second aliquot (10 uL) is added
- 6أخرىThe unreacted biotin is removed by size exclusion chromatography
الإجراء التجريبي
About 2.0 mg of DBP is buffer exchanged with 20 mM bicarbonate at pH 9.6 three times to give a final volume of 1.0 mL. Two aliquots of sulfo-NHS-LC-biotin (Pierce) are dissolved in water to give a 1 mg/mL solution just prior to addition. The first aliquot (10 uL) is added and the reaction is maintained at room temperature for 5 minutes. The second aliquot (10 uL) is added and the reaction proceeds for an additional 4 minutes. The unreacted biotin is removed by size exclusion chromatography using SEPHADEX G-25 and 100 mM PBS, pH 7.4 as the mobile phase.